Rapid detection of Salmonella enterica in primary production samples by eliminating DNA amplification inhibitors using an improved sample pre-treatment method.

Sensitive detection of pathogens in livestock farms is an integral part of the One Health Action Plan of the European Union (EU). Ensuring this requires on-site testing devices that are compatible with complex matrices such as primary production samples. Among all, faeces are considered the most challenging matrix type that makes it difficult to identify pathogens because of complexity in sample preparation for molecular testing. We have developed a loop-mediated isothermal amplification (LAMP) based veterinary point-of-care (POC) device (VETPOD) and adapted it to detect Salmonella enterica in primary production samples. Three different sampling methods (semi-wet chicken faeces, boot socks collection and dust samples from poultry shed) were iteratively tested to assess their nature of complexity and possibility for adapting them as suitable sampling methods for on-site testing. During the study, the sample preparation method that included a two-step centrifugation combined with washing of the enriched Salmonella cells was found crucial in eliminating amplification inhibitors originating from the faecal matrices. A total of 90 samples were tested that included 60 samples for sensitivity study and 30 samples for relative level of detection (RLOD, a level of detection in comparison to ISO 6579:1 reference method). Overall, the VETPOD had a sensitivity of 90%, 84.62% and 81.82% for boot sock, faecal and dust samples, respectively. The RLOD was 2.23 CFU/25 g which was found to be 1.33 times higher than the ISO 6579:1. Performing with an excellent agreement with ISO 6579:1, the VETPOD proved as a promising alternative to detect Salmonella spp. in primary production and animal husbandry samples.

Validation of Point-of-Care Device for Rapid Detection of Salmonella enterica in Meat Products.

Accurate and rapid detection of pathogens in foods of animal origin has been a critical part of the One Health Action Plan of the European Union (EU). Biosensors have the potential in bringing required technologies to accomplish this on the field, wherein loop-mediated isothermal amplification (LAMP) and lab-on-a-chip have proven to be ideal. We have developed a LAMP-based point-of-care (POC) device, the VETPOD, as a solution to the contemporary challenges in the rapid detection of Salmonella spp. The core technology in the VETPOD is a ready-to-use cartridge that included an injection-molded polymer chip with pyramid-shaped optical structures embedded within the chip. These pyramid-shaped optical structures direct the incident light, due to total internal reflection (TIR), through the reaction chambers to the phototransistor. The VETPOD was validated against the ISO 6579-1 reference method. A total of 310 samples were tested that included 180 Salmonella spiked samples in 6 different meat categories and 130 strains to determine the specificity. The overall results were satisfactory, wherein the VETPOD had an acceptable sensitivity (96.51%) compared to the reference (98.81%) and near perfect agreement with ISO 6579-1 with an overall Cohen's kappa of 0.94. The relative level of detection (RLOD) for the VETPOD was 1.38 CFU/25 g that was found to be 1.17 times higher than the reference. The VETPOD showed 98% precision for inclusivity and 100% precision for the exclusivity samples. The VETPOD proved as a useful alternative to detect Salmonella spp. that can be adaptable to a broader spectrum of pathogens in future.

PATHPOD - A loop-mediated isothermal amplification (LAMP)-based point-of-care system for rapid clinical detection of SARS-CoV-2 in hospitals in Denmark.

Sensitive and rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a vital goal in the ongoing COVID-19 pandemic. We present in this comprehensive work, for the first time, detailed fabrication and clinical validation of a point of care (PoC) device for rapid, onsite detection of SARS-CoV-2 using a real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) reaction on a polymer cartridge. The PoC system, namely PATHPOD, consisting of a standalone device (weight less than 1.2 kg) and a cartridge, can perform the detection of 10 different samples and two controls in less than 50 min, which is much more rapid than the golden standard real-time reverse-transcription Polymerase Chain Reaction (RT-PCR), typically taking 16-48 h. The novel total internal reflection (TIR) scheme and the reactions inside the cartridge in the PoC device allow monitoring of the diagnostic results in real-time and onsite. The analytical sensitivity and specificity of the PoC test are comparable with the current RT-PCR, with a limit of detection (LOD) down to 30-50 viral genome copies. The robustness of the PATHPOD PoC system has been confirmed by analyzing 398 clinical samples initially examined in two hospitals in Denmark. The clinical sensitivity and specificity of these tests are discussed.

Natural Rubber-TiO2 Nanocomposite Film for Triboelectric Nanogenerator Application.

In this research, natural rubber (NR)-TiO2 nanocomposites were developed for triboelectric nanogenerator (TENG) application to harvest mechanical energy into electrical energy. Rutile TiO2 nanoparticles were used as fillers in NR material to improve dielectric properties so as to enhance the energy conversion performance of the NR composite TENG. The effect of filler concentration on TENG performance of the NR-TiO2 composites was investigated. In addition, ball-milling method was employed to reduce the agglomeration of TiO2 nanoparticles in order to improve their dispersion in the NR film. It was found that the TENG performance was significantly enhanced due to the increased dielectric constant of the NR-TiO2 composite films fabricated from the ball-milled TiO2. The TENG, fabricated from the NR-TiO2 composite using 24 h ball-milled TiO2 at 0.5%wt, delivered the highest power density of 237 mW/m2, which was almost four times higher than that of pristine NR TENG. Furthermore, the applications of the fabricated NR-TiO2 TENG as a power source to operate portable electronics devices were also demonstrated.

Fabrication of a three-terminal graphene nanoelectromechanical switch using two-dimensional materials.

An alternative three-terminal (3T) subthermal subthreshold slope (SS) switch is required to overcome the exponential increase in leakage current with an increase in the drive current of CMOS devices. In this study, we present a 3T graphene nanoelectromechanical (3T-GNEM) switch with a physically isolated channel in the off-state by using heterogeneously stacked two-dimensional (2D) materials. Hexagonal boron nitride (h-BN) was used as a dielectric layer, and graphene was used as a the top double-clamped beam drain, gate and source electrode material; the drain, gate, and source layers were stacked vertically to achieve a small footprint. The drain to source contact is normally open with an air gap in the off-state, and the gate voltage is applied to mechanically deflect the drain terminal of the doubly clamped graphene beam to make electric contact with the source terminal for the on-state. This 3T-GNEM switch exhibits an SS as small as 10.4 mV dec-1 at room temperature, a pull-in voltage less than 6 V, and a switching voltage window of under 2 V. Since the source and drain terminals are not connected physically in the off-state, this 3T-GNEM switch is a promising candidate for future high-performance low-power logic circuits and all-2D flexible electronics.

Large-area single-crystal graphene grown on a recrystallized Cu(111) surface by using a hole-pocket method.

We describe an efficient chemical vapor deposition (CVD) method for synthesizing graphene with a single crystal orientation on the whole surface of a copper (Cu) foil. We specifically synthesized graphene on the inner surface of a folded Cu foil, on which small holes were made for regulating the permeation and adsorption of the gases used for the synthesis. We compared the results of this method, which we call a "hole-pocket" method, with previously developed methods involving traditional synthesis on an open Cu foil and a Cu "pita-pocket". From these comparisons, we found the orientation of recrystallized Cu to depend on the shape of the Cu foil. Our hole-pocket method did not require treatment of the Cu surface with a complicated process to reduce the density of nucleation seeds in order to synthesize large hexagonal graphene grains, nor did it require the use of a single-crystalline substrate because methane passing through holes on the upper side of the hole-pocket slowly decomposed into carbon atoms and the control of the evaporation of Cu inside the foil pocket helped induce a transformation of the Cu domains to Cu(111). The current hole-pocket method resulted in domains that were both large, ranging from 2-5 mm in size, and oriented in the same manner. By extending the synthesis time, we were able to obtain continuous large-area films of single-crystalline orientation on the whole surface with dimensions of several centimeters.